Stabilized edible oil and fat compositions containing oil of tempeh

ABSTRACT

Tempeh and components thereof, such as oil of tempeh, an extract of tempeh, are useful as stabilizers for food compositions, particularly edible fats and oils. An edible fat or oil containing tempeh or oil of tempeh or a component of oil of tempeh exhibits improved anti-oxidative properties and stability.

United States Patent 1191 Gyorgy 1 1 Oct. 2, 1973 1 1 STABILIZED EDIBLEOIL AND FAT COMPOSITIONS CONTAINING OIL OF TEMPEH [76] Inventor: PaulGyorgy, 201 Curwen Rd.,

Rosemont, Pa. 19010 [22] Filed: June 21, 1971 [21] Appl. No.: 155,252

52 11.5. C1. 99/163, 99/118 151 1111.0 ..A23d 5/04 58 Field of Search99/163, 9, 17, 118 R,

99/118 11,119,122 R, 122 M, 123, 164', 260/112 R, 123.5; 195/4, 29 R, 31R [56] References Cited UNITED STATES PATENTS 2,176,030 10/1939 Musher99/163 2.694645 11/1954 Gleim 99/163 l/1966 Hesseltine et al.... 99/9 X3/1966 Hesseltine et a1 99/9 X OTHER PUBLICATIONS Chemical Abstracts,Volume 61-paragraph 13808 (b),

1964, Teruo Ota et al.

Chemical Abstracts, Volume 69-paragraph 42538 (m), 1968, H. lkehata etal.

Chemical Abstracts, Volume SEE-paragraph 9559 (h), 1962, K. H.Steinkraus et al. I

P. Gyorgy et al., Nature (London) 203, pp. 870872, Aug. 22, 1964.

Primary ExaminerNorman Yucllkoff Assistant Examiner-Kenneth P. Van WyckAtt0rney-Thomas F. Moran et a1.

[57] ABSTRACT 9 Claims, N0 Drawings STABILIZED EDIBLE OIL AND FATCOMPOSITIONS CONTAINING OIL OF TEMPEII This invention relates tomaterials usefuls as stabilizing agents for foods and the like, such asfatty or oily foods, particularly edible oils and fats.

In accordance with one embodiment this invention is directed to foodcompositions, particularly edible oils and fats having improvedstability. In accordance with another embodiment this invention isdirected to the preparation of materials as stabilizers for edible oilsand fats, such as materials useful as antioxidants in food compositions,particularly food compositions containing edible oils and fats. Inaccordance with yet another embodiment this invention is directed to amethod of preparing edible oils and fats and food compositionscontaining the same so as to improve the stability and anti-oxidativeproperties of such compositions. In still yet another embodiment thisinvention is directed to the preparation of materials useful asstabilizing agents in food compositions.

Many food materials, particularly edible fats and oils, i.e. fats andoils of animal and vegetable origin or modified fats and oils ofvegetable and animal origin, become rancid or have an undesirable tasteand/or color imparted thereto during storage, and especially uponexposure to or contact with oxygen.

In accordance with one embodiment it is an object of this invention toprovide edible fat and oil compositions of vegetable or animal originhaving improved stability and anti-oxidative properties.

It is another object of this invention to provide a method of improvingthe anti-oxidative properties of edible fat-or oilcontainingcompositions.

It is another object of this invention to provide a process for themanufacture of anti-oxidative materials, particularly anti-oxidativematerials derived from tempeh, a fermented soybean product.

How these and other objects of this invention are accomplished willbecome apparent in the light of the accompanying disclosure. In at leastone embodiment of the practices of this invention at least one of theforegoing objects will be achieved. a

It has been discovered that tempeh is useful as an antioxidant orstabilizer in food compositions, particularly for edible oils and fats.A component of tempeh which is a fermented soybean product, specificallyoil of tempeh, has been found to be useful as a stabilizer orantioxidant for edible oils and fats. By edible oils and fats is meantthose edible oils and fats of vegetable and- /or animal origin, orchemically modified, e.g., hydrogenated, edible oils and fats ofvegetable and/or animal origin. The practices of this do not includeoils or waxes of mineral or petroleum origin or synthetic hydrocarbonoils and waxes.

In applicants co-pending patent application Ser. No. 32,424 filed Apr.27, 1970 entitled "Food Product Containing Tempeh" itis disclosed thattempeh, particularly lyophilized tempeh, is useful in the preparation offood products, such as fish or fatty meat food products, in that theresulting tempeh-containing food compositions exhibit improvedstability. Applicants aboveidentified co-pending patent application alsodiscloses that during the fermentation of soybeans for the preparationof tempeh, a glycoside of soybean is split with the liberation of anantioxidant which has been identified as 6, 7, 44rihydroxyisoflavone,see P. Gyorgy, K. Murata and H. Ikehata, NATURE (London) 203, 870

(1964). The disclosures of applicants above-identified patentapplication and the above-identified publication are herein incorporatedand made part of this disclosure.

In accordance with one practice of this invention an edible oil or fathaving incorporated therein tempeh, such as dried or lyophilized tempeh,or a component or extract of tempeh, e.g., oil of tempeh, has been foundto possess antioxidative properties.

Oil of tempeh, a component or extract of tempeh, which has been found tobe useful as a stabilizing agent or antioxidant for edible oils and fatsis produced by extracting tempeh, a fermented soybean, with selectivesolvents. For example, oil of tempeh or tempeh oil is produced byextracting tempeh, preferably dry tempeh, obtained by fermentingsoybeans with the fungus Rhizopus oli gosporus, with a mixture of anormally liquid aliphatic hydrocarbon, such as an aliphatic hydrocarboncontaining from 4 to 12 carbon atoms per molecule, e.g., hexane, and anormally liquid oxygencontaining, preferably aliphatic, polar organiccompound containing from 1 to 10 carbon atoms per molecule, such as analkanol, e.g., ethanol. A solvent admixture made up of the aliphatichydrocarbon, e.g. one volume of hexane and a polar oxygen-containingorganic compound, such as an alkanol, e.g., 0.5-6.0 volumes ethanol, isemployed in an amount of about 5 volumes of this solvent admixture toone volume of tempeh. The resulting admixture of solvent and tempeh ismaintained in contact, preferably at about room temperature, e.g. atemperature in the range 1040C., for a sufficient period of time, e.g.in the range from about 2 minutes to about 2 hours, more or less,depending upon the intensity of mixing, temperature, etc. Thereupon, theliquid solvent admixture is separated from the undissolved or extractedtempeh and the separated liquid solvent and oil of tempeh-containingextract phase is then treated, such as by distillation, for the removalof the solvent hydrocarbon and the oxygenated polar organic compoundtherefrom. This remaining material identified herein as oil of tempeh ortempeh oil exhibits remarkable stability and possesses antioxidativeproperties. For example, oil of tempeh when stored for 17 months in afreezer and then for 21 days at a temperature of 37C. exhibited aperoxide value (POV) of about 5.9 meq/kg. In contrast, commercial soyaoil or soybean oil when maintained and tested under the same conditionsexhibited a peroxide value of 81.4.

In these tests and in the other tests demonstrative of the practices ofthis invention the peroxide value was determined according to the methodof D. II. Wheeler Oil & Soap 9, 89 (1932). This method is based upon thefollowing procedure. A sample of the oil to be tested weighing 0.5 gramis placed in a 300 mls. flask with a glass stopper and there are addedthereto 10 mls. of chloroform to dissolve the sample. Thereupon, l5 mls.of glacial acetic acid are added along with about 1 gram of powderedpotassium iodide KI. The resulting admixture is then refluxed on a waterbath for three minutes and then cooled in an ice water bath. Thereuponmls. of distilled water are added and the resulting flask contents werevigorously shaken. After shaking there was added a drop of 1 percentstarch solution and the flask contents then titrated with N/ 1000 sodiumthiosulfate until the iodine 1, color disappeared.

The results of these tests are then calculated on meg/kg.

In accordance with still another embodiment of this invention theabove-described tempeh oil or oil of tempeh, such as is produced bymultiple (5) extraction of 5 dry tempeh with a mixture of two partshexane and one part ethanol is further treated and extracted to producea component thereof exhibiting even more remarkable stabilizing oranti-oxidative properties. This component is obtained by extracting oilof tempeh with a 50 percent aqueous ethanol solution and the solventthen evaporated from the resulting extract phase. After removal ofsubstantially all of the solvent the residue is then extracted withpetroleum ether to produce a liquid oily phase and an insoluble,substantially solid phase. The insoluble phase is separated and 50percent aqueous ethanol added thereto to yield an aqueous phasecontaining the insoluble material dispersed or emulsified therein.Acetone is then added to precipitate the solid material which is thenseparated, such as by filtration, and dried.

Following the above procedure there is recovered on the basis of 1 gramof tempeh oil an anti-oxidatively active component thereof in the amountof about 1 milligram. This material is effective when added on the basisof aboutone part per thousand to an edible oil or fat. such as soybeanoil, to protect the oil or fat for a substantial period of time againstoxidation.

For example, the POV of commercial soybean oil after storage for 8 daysat 37C. was 47. When the soya oil had incorporated therein an amount of0.1 percent by weight of the aforesaid component extracted from tempehoil the resulting soybean oil after storage under similar conditions hada POV of 21.

Further illustrative of the practices of this invention mixtures ofcommercial soybean oil and oil of tempeh prepared in accordance withthis invention were stored at 60C. for a number of days, 35 days and 43days, and the peroxide values of these mixtures then determined. Theresults of these tests are set forth in accompanying Table I:

TABLE I POV After POV After Composition 354 Days 43 Days 100% soybeanoil 118 270 90% soybean oil, 10% tempeh oil 53 82 80% soybean oil, 20%tempeh oil 33 30 70% soybean oil, 30% tempeh oil 25 20 60% soybean oil,40% tempeh oil 10 13 50% soybean oil. 50% tempeh oil 7 8 100% tempeh oil4 5 Further tests were carried out on mixtures of soybean oil and tempehoil to determine stability of mixtures of these oils as measured byperoxide values, after storage for 35 days at 37C. The results of thesetests are set forth in accompanying Table 11:

100% tempeh oil Admixtures of commercial soybean oil and tempeh oil inaccordance with this invention in the amount 40-80 percent by weighttempeh oil and -20 percent by weight soybean oil after storage orincubation for 18 days at 37C. (1 gram in open scintillation countingvials) gave peroxide values in the range 17-5 whereas a commerciallyavailable stabilized soybean oil gave a peroxide value of 23. This testclearly demonstrated the superiority of a soybean oil stabilized withtempeh oil in accordance with this invention over commercially availablestabilized soybean oils.

Further tests were carried out wherein mixtures of corn oil and tempehoil and lard and tempeh oil were stored and incubated for 26 days at atemperature of 38C. (2 grams in open scintillation counting vials) andthe peroxide values then measured. The results of these tests are setforth in accompanhing Table 111:

TABLE III POV After Composition 26 Days 100% lard 220 100% corn oil 9510% tempeh oil. 90% lard 10 10% tempeh oil, 90% corn oil 12 20% tempehoil, lard 5 20% tempeh oil, 80% corn oil 12 40% tempeh oil, 60% lard 640% tempeh oil, 60& corn oil 12 Further illustrative of the advantagesof the practices of this invention the peroxide values of various oilmixtures were determined after storage at 60C. The results of thesetests are set forth in accompanying Table IV:

TABLE IV.POV OF VARIOUS OILS AFTER STORAGE AT 60 C.

Days of storage Oil composition Cottonseed oil 1.9 101.7 202.8 350.6Cottonseed oil 10% tempeh oil 3.3 64.4 113.9 190.7 Cottonseed oil 20%tempeh oil 5.4 34.8 89.2 112.4 Cottonseed oil 30% tempeh oil 3.9 6.626.4 82.5 Cottonseed oil+ 50% tempeh oil 10.8 5.7 5.3 48.3 Tempeh oil16.0 3.9 1.7 5.5

safflower oil 2.5 103.3 323.5 334.3 Safflower oil 10% tempeh oil 3.770.1 133.1 258.4 Safflower oil 20% tempeh oil 6.8 39.6 90.4 168.2Safflower oil 30% tempeh oil 8.6 1 1.6 61.1 100.2

luhlc IV Sal'flower oil 50% tempeh oil Tempeh oil unlmucd Lard Lardtempeh oil Lard 20% tempeh oil Lard 30% tempeh oil Lard 50% tempeh oilTempeh oil Tests (iodine values) were also carried out on two samples oftempeh oil and two samples of commercial soybean oil. The results ofthese tests show that the two samples of tempeh oil gave iodine valuesof 132.6 and 123.5 and the samples of soybean gave iodine values of139.6 and 133.2.

As indicated hereinabove, in the extraction of term peh oil or oil oftempeh from tempeh for use as an antioxidant in accordance with thepractices of this invention various solvents and solvent mixtures mightbe employed. For example, it is preferred to employ as the hydrocarbonsolvent an aliphatic (including cycloaliphatic) hydrocarbon or mixtures,such as a petroleum fraction comprised of predominantly aliphatic(including cycloaliphatic) hydrocarbons. A useful petroleum fractionwould have an initial boiling point in the range 70170F. and an endboiling point in the range 150-350F. Suitable hydrocarbons which mightbe employed in the preparation of solvent admixtures in accordance withthis invention would include the pentanes, e.g., npentane, isopentane,the hexanes, such as n-hexane, and the various isohexanes, the heptanes,such as n-heptane, the octanes, such as n-octane, the nonanes, such asn-nonane. Aliphatic hydrocarbons which would be useful would include thesaturated as well as the unsaturated, especially the monounsaturated orolefinic aliphatic hydrocarbons.

The other solvent, i.e. the oxygen-containing polar aliphatic (includingcycloaliphatic) compound useful would include the various ketones, e.g.acetone, methyl ethyl ketone, methyl isobutyl ketone, as well ascyclohexanone, the ethers, the various alcohols, such as ethanol,butanol, isopropanol as well as cyclohexanol, the various esters, suchas ethyl acetate and the like.

The practices of this invention are broadly applicable to foodcompositions which contain an edible oil or an edible fat. The practicesof this invention are particularly useful for the preparation of edibleoil and edible fats having improved anti-oxidative properties. Edibleoils and fats and compositions, particularly food compositionscontaining edible oils and fats, which are improved by the incorporationtherein of tempeh or oil of tempeh or a component of tempeh prepared inaccordance with your invention would include the animal fats, such astallow, beef fat, pork fat, lard, spermaceti, fats derived from fattyfish, meats and fish food compositions containing fats and the like, theedible oils both of animal and vegetable origin, such as olive oil,cottonseed oil, sperm or whale oil, safflower oil, peanut oil, corn oil,palm oil, cashew nut oil, soybean oil, mixtures thereof and thechemically modified, e.g. hydrogenated, fats and oils.

Other food compositions which have tempeh or oil of tempeh or acomponent of oil of tempeh usefully incorporated therein include specialfood products, particularly ready made or ready to use food products,for instance, soya meal or flour, milk powder and grain or crackers.

Substantial benefits of the practices of this invention are obtainablewhen only a minor amount of tempeh is incorporated in food compositions,particularly fat or oil-containing food compositions. Usually an amountupwards of about 0.5% of tempeh or oil of tempeh substantially improvesthe anti-oxidative properties of compositions containing the same. It ispreferred in accordance with the practices of this invention toincorporate tempeh oil and tempeh or a component or extract of oil oftempeh in food compositions in an amount in the range from about 1percent by weight up to about 20-25 percent by weight, more or less.

As will be apparent to those skilled in the art in the light of theforegoing disclosure, many modifications, alterations and substitutionsare possible in the practice of this invention without departing fromthe spirit or scope thereof.

1 claim:

liquid extract phase and separating therefrom the admixture of saidaliphatic hydrocarbon and said polar organic compound to produce saidoil of tempeh.

2. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is animal fat.

3. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is a vegetable oil.

4. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is a polyunsaturated oil.

5. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is corn oil.

6. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is cottonseed oil.

7. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is lard.

8. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is olive oil.

9. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is soybean oil.

2. A stabilized edible composition in accordance with claim 1 whereinsaid edible fat or oil is animal fat.
 3. A stabilized edible compositionin accordance with claim 1 wherein said edible fat or oil is a vegetableoil.
 4. A stabilized edible composition in accordance with claim 1wherein said edible fat or oil is a polyunsaturated oil.
 5. A stabilizededible composition in accordance with claim 1 wherein said edible fat oroil is corn oil.
 6. A stabilized edible composition in accordance withclaim 1 wherein said edible fat or oil is cottonseed oil.
 7. Astabilized edible composition in accordance with claim 1 wherein saidedible fat or oil is lard.
 8. A stabilized edible composition inaccordance with claim 1 wherein said edible fat or oil is olive oil. 9.A stabilized edible composition in accordance with claim 1 wherein saidedible fat or oil is soybean oil.